[0001] The present invention relates to the use of substrates of an UDP-glucuronosyltransferase
(UGT) and salts thereof, for the prevention and /or the treatment of neurological
disorders. In preferred embodiment, the present invention relates to the use of substrates
of at least an UDP-glucuronosyltransferase (UGT) expressed in brain and salts thereof,
for the prevention and /or the treatment of neurological disorders. It further relates
to the use of said substrates, and salts thereof, for preventing and/or treating glutamate
cytotoxicity, and more specifically of glutamate induced neurological disorders. Additionally,
it concerns the use of said substrates, and salts thereof, for making drugs exerting
an inhibitory effect on the release of glutamate.
[0002] A large number of studies have established that cellular communication using excitatory
amino acids can be transformed into a mechanism of cell destruction.
[0003] Glutamate, for example, is the main excitatory neurotransmitter in the nervous system,
especially brain and spinal cord, of mammals wherein it is working at a variety of
excitatory synapses.
[0004] The ubiquitous distribution of glutamate receptors throughout the nervous system
proves that glutamate plays a central role in a wide range of physiological as well
as pathological events (Watkins J. C., Collingridge G. L., The NMDA receptor, IRL
Oxford, 1989). It is for example strongly suggested that it plays a central role in
functions such as learning, pattern recognition, and memory (Bliss T. V. P. Collingridge
G. L., Nature 361, 31-39, 1993).
[0005] Normally extracellular levels of glutamate are elevated only in a brief and spatially
localized fashion associated with normal synaptic transmission; however, under pathologic
circumstances levels may remain dramatically increased.
[0006] Additionally, it has also been known for decades that glutamate is toxic to neurons
in vitro and
in vivo and that the function of glutamate receptors, especially glutamate receptors of the
N-methyl-D-aspartate ("NMDA") receptor subtype, is crucial in a number of neuronal
damages and injuries (Appel S. H., Trends Neurosci. 16, 3-5, 1993). Many neurological
disorders involving epileptic seizures and chronic or acute degenerative processes,
such as for example Alzheimer's, Huntington's, Parkinson's diseases, multiple sclerosis
(MS), amyotrophic lateral sclerosis (ALS), spinal muscular atrophy (SMA), retinopathy,
stroke, depression and traumatic brain injury, involve neuronal cell death caused
by over-stimulation of the glutamate receptors. Similarly, it has been shown that
neuronal injury caused by ischemia after occlusion of cerebral arteries could, at
least partially, be mediated by excessive activation of glutamate receptors as in
the ischemic brain, extracellular glutamate is elevated rapidly after the onset of
ischemia and declines following reperfusion (Davalos et al., 1997, Stroke, 28, 708-710).
Other pathologic circumstances associated with dramatic increase of extracellular
glutamate levels are hypoxia or hypoglycaemia. Finally, Stephans and Yamamoto (1994,
Synapse, 17, 203-209) have shown that drug-induced neurotoxicity, for example neurotoxic
effects of methamphetamine (METH) on striatal dopaminergic neurons, could actually
be mediated by over-stimulation of the glutamate receptors.
[0007] These excessive activations of glutamate receptors, referred to as "glutamate cytotoxicity",
are actually associated with the elevation of extracellular glutamate levels. The
mechanisms of the elevation of extracellular glutamate include enhanced efflux of
glutamate and/or the reduction of glutamate uptake by cells. Thus, it would be desirable
to provide a means of protecting affected cells, especially neurons, from glutamate-induced
cytotoxicity, and more specifically to provide means of regulating glutamate release
and/or uptake by glutamate producing cells.
[0008] To this end, it has already been proposed to target the glutamate receptors, mostly
the N-methyl-D-aspartate ("NMDA") receptor, present on the targeted cells by inhibiting
them by the use of agonist or antagonist specific molecules. Examples of such molecules
are anthranilic acid derivatives (see US 5,789,444), Basilen Blue D-3G (Reactive Blue
2) and Cibacron Blue 3GA and 5-adenylylimidodiphosphate (AMPPNP) (see US 6,326,370),
NMDA specific antagonists such as ketamine, gluconate (Sakaguchi et al., 1999, Neuroscience,
92, 677-684), dextromophan, or 3-(2-carboxypiperazin-4-yl)-propyl-1-phosphonic acid
(Kristensen et al., 1992, Pain, 51:249-253; Eide et al., 1995, Pain, 61,221-228),
or the 2-methyl-6-(phenylethynyl)pyridine (MPEP) which is an antagonist of the metabotropic
glutamate receptor subtype 5 (mGluR5) (Ossowska et al., 2001, Neuropharmacology, 41,
413-420).
[0009] However, widespread use of these compounds is precluded by their undesirable side
effects (e.g. psychotomimetic effects, headache, hallucinations, dysphoria or disturbances
of cognitive and motor functions). Thus, the available treatment methods are not satisfactory
in terms of safety or efficiency for their wide implementation.
[0010] Therefore, there is still a need in the provision of improved methods and means for
protecting affected cells, and more preferably neurons, from glutamate-induced cytotoxicity.
[0011] Similarly, many neurological disorders are associated with elevated levels of neuromediator
(or neurotransmiter) release such as for example acetylcholine (Ach), butyrylcholine
(BuCh), gamma-aminobutyrate (GABA), serotonin or the catecholamines, in particular
dopamine or ATP.
[0012] In mammals, glucuronidation represents a major metabolic pathway which enhances the
detoxification and elimination of many lipophilic drugs, pollutants, xenobiotics and
endogenous compounds by converting them in less biologically available, and thus less
active, glucuronidated substances exhibiting higher polarity and water-solubility,
thus facilitating their transport to excretory organs and subsequent excretion in
urine or bile (for a review, see Ritter, 2000, Chemico-Biological Interactions, 129,
171-193). The reaction is catalyzed by UDP-glucuronosyltransferases (UGTs). This multigenic
family of enzymes transfer the glucuronic acid moiety of UDP-glucuronic acid to structurally
unrelated compounds through hydroxyl (alcoholic, phenolic), carboxyl, sulfuryl, carbonyl
and amino (primary, secondary or tertiary) linkages (for review, see Tukey and Strassburg,
2000, Annu. Rev. Pharmacol., Toxicol., 40, 581-616). Initially, glucuronidation has
been considered to represent a metabolic pathway performed mainly in the liver. However,
multiple studies have further indicated that UGT activity was also present in human
intestinal, kidney, colon tissue and brain where this enzyme is believed to take part
actively in the defence of the organism against potentially reactive hydroxylated
molecules which can reach the organ (Suleman et al., 1998, Archives of Biochemistry
and Biophysics, 358, 1, 63-67 ; Gradinaru et al., 1999, Neurochemical Research, 24,
995-1000).
[0013] The inventors have now shown that the UGT activity
in vivo, and more specifically in brain, can be turned away and used for defining a new class
of compounds for the treatment and/or prevention of acute and chronic neuromediator-related
diseases or conditions, particularly neurological diseases. The present invention
is a pharmacological alternative to previously described methods implementing compounds,
such as competitive and non-competitive glutamate antagonists or agonists, gangliosides
and growth factors. In special embodiments, the present invention provides a new class
of compounds which can be used as pharmacological tools for the modulation of neuromediator
cellular release and cytotoxicity, preferably neurotoxicity, and which allows the
possible treatment and/or prevention of many neurological disorders involving epileptic
seizures and acute and chronic neurodegenerative diseases, as well as neuronal injury
caused by ischemia or neuromediator-related diseases or conditions, wherein said disorders
are, at least partially, associated with excessive activation of neuromediator receptors
and/or with excessive extracellular neuromediator levels.
[0014] First, the invention concerns the use of at least one substrate of an UDP-glucuronosyltransferase
(UGT), and salts thereof, for the preparation of a pharmaceutical composition having
an inhibitory effect on the extracellular neuromediator release into an individual
treated with said composition. According to one special embodiment, said UDP-glucuronosyltransferase
(UGT) is expressed in brain.
[0015] According to specific embodiment, said UDP-glucuronosyltransferase (UGT) is selected
among the group consisting of UDP-glucuronosyltransferase 1A1 (UGT1A1, also termed
HUG-Br1 or UGT1.1), UDP-glucuronosyltransferase 1A3(UGT1A3, also termed 1c), UDP-glucuronosyltransferase
1A4 (UGT1A4, also termed HUG-Br2), UDP-glucuronosyltransferase 1A6 (UGT1A6), UDP-glucuronosyltransferase
1A9 (UGT1A9, also termed HLUG P4), UDP-glucuronosyltransferase 2B4 (UGT2B4, also termed
Hlug-25, Th-1, h-1, or h-20), UDP-glucuronosyltransferase 2B7 (UGT2B7), UDP-glucuronosyltransferase
2B10 (UGT2B10, also termed h-46), UDP-glucuronosyltransferase 2B15 (UGT2B15, also
termed UDPGTh-3,h-3, HLUG-4, or HE 8A), UDP-glucuronosyltransferase 2B17 (UGT2B17)
and UDP-glucuronosyltransferase 2B28 (UGT2B28). According to specific embodiment,
said UDP-glucuronosyltransferase (UGT) is expressed in brain and is selected among
the group consisting of UDP-glucuronosyltransferase 1A6 (UGT1A6) and UDP-glucuronosyltransferase
2B7 (UGT2B7).
[0016] According to the present invention, the term "UDP-glucuronosyltransferase (UGT)"
refers to enzymes catalyzing the glucuronidation reaction. Said reaction utilizes
UDP-glucuronic acid as a cosubstrate for the formation of glucuronides. All of these
terms are widely used in the litterature; for a review, please refer to Tukey and
Strassburg, 2000, Annu. Rev. Pharmacol. Toxicol., 40, 581-616. The method of choice
for analysing the activity of a define UGT is the expression of the corresponding
cDNA in a cell line that express minimal levels of the respective protein. More precisely,
cells are transfected by standard transient transfection methods or by stable introduction
of the cDNA into the genome following selection of the cells with an appropriate antibiotic.
Then the cells are homogenized in a buffer containing Tris-HCl and MgCl
2 and subjected to sonication. Cell homogenates are then tested for their ability to
glucuronidate series of substrates. The activity and selectivity of the UGT for a
substrate could be compared with known specific substrates if available (Remmel and
Burchell, 1993, Biochem. Pharmacol., 46, 559-566).
[0017] Currently, 15 UGT cDNAs have been identified in human, eight UGT1A proteins encoded
by the UGT1A locus and seven proteins encoded by UGT2B genes. "UDP-glucuronosyltransferase
(UGT) 1A6" refers to one of the UGT proteins encoded by the UGT1A locus. The human
UGT1A6 sequence is presented in SEQ ID NO:1 (Harding et al., 1988, Proc. Natl. Acad.
Sci. 85, 8381-8385 ). UGT1A6 is also called PNP-UGT, phenol p I 6.2, UGT1*6, UGT1A06
or HLUGP1. "UDP-glucuronosyltransferase (UGT) 2B7" refers to one of the UGT proteins
encoded by the UGT2B locus. The human UGT2B7 sequence is presented in SEQ ID NO:2
(Ritter et al., 1990, J. Biol. Chem., 265, 7900-7906. UGT2B7 is also named Th-2, h-2,
Hlug-6, UGT2*7 or Catechol Estrogen UDPGT.
[0018] According to the present invention the term "UGT substrate" generally designates
both the aglycone substrate (i.e. compound which can be enzymatically converted, or
metabolized, by the UGT enzyme as disclosed above) and the glucuronidated substrate
(i.e. compound which has been converted by addition of at least one glucuronic acid
moiety through active groups).
[0019] According to the present invention the term "neuromediator", a synonym of "neurotransmitter",
is intended to designate chemicals that transmit information across the junction (synapse)
that separates one nerve cell (neuron) from another nerve cell or a muscle. Examples
of said neuromediators are acetylcholine (Ach), butyrylcholine (BuCh), gamma-aminobutyrate
(GABA), serotonin, norepinephrine, epinephrine, endorphins, or the catecholamines,
in particular dopamine or adenosine triphosphate. According to one preferred embodiment,
said neuromediator is the glutamate.
[0020] According to one embodiment, the UGT substrate of the invention is selected among
the group consisting of planar and small phenols, polycyclic aromatic hydrocarbons,
and compounds which are structurally related. More specifically, said UGT substrate
contains at least one moiety selected in the group consisting in hydroxyl (alcoholic,
phenolic, etc...), carboxyl, sulfuryl, carbonyl and amino (primary, secondary or tertiary)
moieties.
[0021] According to a specific embodiment, said substrate is a compound which can be converted
by at least the UDP-glucuronosyltransferase 1A6 (UGT1A6) and is selected in the group
consisting of 1-Naphthol, 2-Naphthol , 4-nitrophenol , methylsalicylate, ketoprofen
, naproxen, 5-OH tryptamine/serotonin, carprofen/rimadyl , acetaminophen/paracetamol
,benzidine, 4-methylumbelliferone (4-MU), silymarin (Venketaramanan et al., 2000,
Drug Metab Dispos., 28,1270-3 ; silymarin is a mixture of toxifolin, silichristin,
silidianin, silybin A et B, isosilybin A et B) [see Figure 1A and 1B].
[0022] According to another embodiment, said substrate is a compound which can be converted
by at least the UDP-glucuronosyltransferase 2B7 (UGT2B7) and is selected in the group
consisting of transretinoic acid, ASA , AZT , benoxaprofen , benzidine, (benzo(a)pyrene
mbs), buprenorphine, carprofen, chloramphenicol, clofibric acid flavenoids quercetin,
kaempfenol, cyclosporin, DMXAA, diclofenac, dihydrocodeine DHC, dihydromorphone ,
mefenamic acid, fenemate NSAID, mycophenolic acid MCPA mofetil, fenoprofen, hydromorphone,
ibuprofen, ketoprofen, linoleic acid , lorazepam, losartan, menthol, morphine 3 and
morphine 6, nalbufene, nalmefene, naltrindole, nalorphine, naloxone, naltrexone, S-naproxen,
norcodeine, normorphone, oxycodone, oxymorphone, pirprofen, propanolol, S-oxazepam,
tacrolimus, temazepam, tolcapone, tiaprofenic, valproate, zomepirac, 5-OH tryptamine.
[0023] According to a specific embodiment, said substrate is selected in the group consisting
of ketoprofen , naproxen, 5-OH tryptamine/serotonin, carprofen/rimadyl and benzidine.
[0024] In special embodiments, the UGT substrates of the invention are further substituted
with, one to four, identical or different, heteroatoms and/or hetero groups. Examples
of said heteroatoms and/or hetero groups are O, H, alkyl or aryl groups C
nH
n+1, with n = 1 to 5, OCH
3, N, halogens (fluorine, chlorine, bromine, or iodine atom), S or any labeling element
allowing to visualize said substrates. These substituting atoms or groups, and their
uses, are widely known in the art.
[0025] The addition salts of the substrates of the invention comprise conventional salt
formed from inorganic or organic acids or bases, such as hydrochloric, hydrobromic,
sulfuric, phosphoric, nitric, perchloric, fumaric, acetic, sodium, lithium, potassium,
magnesium, aluminium, calcium, zinc, ethylenediamine ; formic, benzoic, maleic, tartaric,
citric, oxalic, aspartic acid, and alkane-sulfonic acids is even mentioned.
[0026] The UGT substrates used according to the invention are commercially available in
many commercial catalogues for example in Merck, "Produits chimiques et réactifs"
2002 and Acros Organics "Fine Chemicals" 2002-2003 :
1-Naphtol : |
822289 (Merck) |
2-Naphtol : |
822290 (Merck) |
4-Nitrophenol : |
820896 (Merck) |
5OH Tryptamine/serotonin |
21502-1000 (Acros) |
4-methylumbelliferone |
40549-0010 (Acros) |
All trans retinoic acid |
20734-1000 (Acros) |
Chloramphenicol |
22792-0250 (Acros) |
Clofibric acid |
18909-0500 (Acros) |
[0027] Further suppliers are :
- Naproxen :
- Apranax (FR, Roche) or Naprosyn (US, Roche)
- Carprofen :
- Rimadyl (Pfizer)
- Ketoprofen :
- Toprec (FR, Aventis) or Orudis (US, Wyeth).
[0028] The newly identified inhibitory properties of these UGT substrates make them particularly
suitable for treating and/or preventing diseases, conditions and attacks related to
deleterious effects of neurotransmitter released, and preferably neurological ones.
In one special embodiment, the UGT substrates of the invention are suitable for treating
and/or preventing diseases, conditions and attacks related to deleterious effects
of glutamate released, and preferably neurological ones.
[0029] According to one special embodiment, said diseases, conditions and attacks are related
to deleterious effects of neurotransmitter released in excess, and more particularly
to deleterious effects of glutamate released in excess.
[0030] Thus the present invention further relates to a method for treating and/or preventing
neuromediator-evoked cytotoxicity in a patient in need thereof comprising administering
to said patient a composition containing a therapeutically effective amount of at
least one substrate of an UDP-glucuronosyltransferase (UGT) as defined above and a
pharmaceutically acceptable carrier. In special embodiment, said UGT substrate is
a compound which can be converted by at least one UDP-glucuronosyltransferase (UGT),
and preferably by at least one UDP-glucuronosyltransferase (UGT) expressed in brain.
[0031] Preferably, the present invention relates to a method for treating and/or preventing
glutamate-evoked cytotoxicity in a patient in need thereof comprising administering
to said patient a composition containing a therapeutically effective amount of at
least one substrate of an UDP-glucuronosyltransferase (UGT) as defined above and a
pharmaceutically acceptable carrier. In special embodiment, said UGT substrate is
a compound which can be converted by at least one UDP-glucuronosyltransferase (UGT),
and preferably by at least one UDP-glucuronosyltransferase (UGT) expressed in brain.
[0032] According to another embodiment, said UGT substrate is selected among the group consisting
of planar and small phenols, polycyclic aromatic hydrocarbons, and compounds which
are structurally related. More specifically, said UGT substrate contains at least
one moiety selected in the group consisting in hydroxyl (alcoholic, phenolic, etc...),
carboxyl, sulfuryl, carbonyl and amino (primary, secondary or tertiary) moieties.
[0033] According to another embodiment, said substrate is a compound which can be converted
by the UDP-glucuronosyltransferase 1A6 (UGT1A6) and is selected in the group consisting
of 1-Naphthol, 2-Naphthol , 4-nitrophenol , methylsalicylate, ketoprofen , naproxen,
5-OH tryptamine/serotonin, carprofen/rimadyl , acetaminophen/paracetamol , benzidine,
4-methylumbelliferone (4-MU), silymarin (Venketaramanan et al., 2000, Drug Metab Dispos.,
28,1270-3 ; Silymarin is a mixture of toxifolin, silichristin, silidianin, silybin
A et B, isosilybin A et B) [see Figure 1].
[0034] According to another embodiment, said substrate is a compound which can be converted
by the UDP-glucuronosyltransferase 2B7 (UGT2B7) and is selected in the group consisting
of transretinoic acid, ASA , AZT , benoxaprofen, benzidine, (benzo(a)pyrene mbs),
buprenorphine, carprofen, chloramphenicol, clofibric acid , flavenoids quercetin,
kaempfenol, cyclosporin, DMXAA, diclofenac, dihydrocodeine DHC, dihydromorphone, mefenamic
acid, fenemate NSAID, mycophenolic acid MCPA mofetil, fenoprofen, hydromorphone, ibuprofen,
ketoprofen, linoleic acid , lorazepam, losartan, menthol, morphine 3 and morphine
6, nalbufene, nalmefene, naltrindole, nalorphine, naloxone, naltrexone, S-naproxen,
norcodeine, normorphone, oxycodone, oxymorphone, pirprofen, propanolol, S-oxazepam,
tacrolimus, temazepam, tolcapone, tiaprofenic, valproate, zomepirac, 5-OH tryptamine.
[0035] According to a special embodiment, said substrate is selected in the group consisting
of ketoprofen, naproxen, 5-OH tryptamine/serotonin, carprofen/rimadyl and benzidine.
[0036] The term "neuromediator-evoked cytotoxicity" or neurotransmitter-evoked cytotoxicity"
within the present invention is intended to designate cell toxicity associated with
excessive activations of the concerned neuromediator receptors. These terms are well
known by the one skilled in the art. More specifically, the "glutamate-evoked cytotoxicity"
concerns all affected cells expressing glutamate receptors. According to preferred
embodiments, these cells are susceptible to be affected by neuromediators are nervous
cells (i.e. neuro-cells), preferably neurons. These affected nervous cells are, for
example, present in brain, spinal cord, retina, at the neuro-muscular junction, etc
... "Cytotoxicity" means that the cell functions and/or properties are affected, leading
to cell malfunctioning, and finally to cell death.
[0037] In a particularly preferred embodiment, the method of the invention is intended for
treating and/or preventing neuromediator-evoked neurotoxicity, and even more preferably
for treating and/or preventing neurodegeneration (i.e. degeneration of nervous cells).
[0038] In another particularly preferred embodiment, the method of the invention is intended
for treating and/or preventing glutamate-evoked neurotoxicity, and even more preferably
for treating and/or preventing neurodegeneration (i.e. degeneration of nervous cells).
[0039] The present invention further relates to a method for modulating the release of at
least one neuromediator in a patient comprising administering to said patient a composition
containing a therapeutically effective amount of at least one substrate of an UDP-glucuronosyltransferase
(UGT) and a pharmaceutically acceptable carrier, wherein said substrate is as detailed
above. In special embodiment, said UGT substrate is a compound which can be converted
by at least one UDP-glucuronosyltransferase (UGT), and preferably by at least one
UDP-glucuronosyltransferase (UGT) expressed in brain.
[0040] The present invention further relates to a method for modulating the release of glutamate
in a patient comprising administering to said patient a composition containing a therapeutically
effective amount of at least one substrate of an UDP-glucuronosyltransferase (UGT)
and a pharmaceutically acceptable carrier, wherein said substrate is as detailed above.
In special embodiment, said UGT substrate is a compound which can be converted by
at least one UDP-glucuronosyltransferase (UGT), and preferably by at least one UDP-glucuronosyltransferase
(UGT) expressed in brain.
[0041] "Modulating the release of neuromediator" means that the levels of released neuromediator
in non treated patient is different from the one observed after his treatment with
the substrates of the invention. According to one embodiment, treatment of the patient
with the substrates of the invention leads to a negative modulation, preferably to
the inhibition, of the neuromediator release by the producing cells, and thus to a
decreased neuromediator level in the treated patient compared to the same neuromediator
level observed before said treatment.
[0042] The present invention further relates to a method for treating and/or preventing
disease and/or condition associated with the excessive release of at least one neuromediator
in a patient comprising administration to said patient of a composition containing
a therapeutically effective amount of at least one substrate of an UDP-glucuronosyltransferase
(UGT) and a pharmaceutically acceptable carrier, wherein said substrate is as detailed
above. In special embodiment, said UGT substrate is a compound which can be converted
by at least one UDP-glucuronosyltransferase (UGT), and preferably by at least one
UDP-glucuronosyltransferase (UGT) expressed in brain.
[0043] The present invention further relates to a method for treating and/or preventing
disease and/or condition associated with the excessive release of glutamate in a patient
comprising administration to said patient of a composition containing a therapeutically
effective amount of at least one substrate of an UDP-glucuronosyltransferase (UGT)
and a pharmaceutically acceptable carrier, wherein said substrate is as detailed above.
In special embodiment, said UGT substrate is a compound which can be converted by
at least one UDP-glucuronosyltransferase (UGT), and preferably by at least one UDP-glucuronosyltransferase
(UGT) expressed in brain.
[0044] According to the invention, the terms "treating and/or preventing" refer to a process
that is intended to produce a beneficial change in the condition of a mammal, e.g.,
a human, often referred to as a patient. A beneficial change can, for example, include
one or more of: restoration of function, reduction of symptoms, limitation or retardation
of progression of a disease, disorder, or condition or prevention, limitation or retardation
of deterioration of a patient's condition, disease or disorder. Such therapy can involve,
for example, nutritional modifications, administration of radiation, administration
of a drug, behavioural modifications, and combinations of these, among others.
[0045] According to the invention, "disease and/or condition associated with the excessive
release of at least one neuromediator" is intended to designate large number of acute
and chronic neuromediator-related diseases or conditions, particularly neurological
diseases. It designates more specifically epileptic seizures and acute and chronic
neurodegenerative diseases, as well as neuronal injury caused by ischemia or neuromediator-related
diseases or conditions, wherein said disorders are, at least partially, associated
with excessive activation of neuromediator receptors and/or with excessive extracellular
neuromediator levels. Examples are involving chronic or acute degenerative disorders,
such as for example Alzheimer's, Huntington's, Parkinson's diseases, multiple sclerosis
(MS), amyotrophic lateral sclerosis (ALS), spinal muscular atrophy (SMA), retinopathy,
stroke, clinical depression and traumatic brain injury, involve neuronal cell death
caused by over-stimulation of the neuromediator receptors, and more specially of the
glutamate receptors. Similarly, neuronal injury caused by ischemia or drug-induced
neurotoxicity, for example neurotoxic effects of methamphetamine (METH) on striatal
dopaminergic neurons, are indications of the methods according to the present invention.
Other indications are the neuromediator-related conditions such as for example pain,
hormonal balance, blood pressure, thermoregulation, respiration, learning, pattern
recognition or memory, or any disorder subsequent to hypoxia or hypoglycaemia, especially
when these indications are glutamate-related conditions.
[0046] For example, the treatment of epilepsy, clinical depression, amyotrophic lateral
sclerosis, spinal muscular atrophy (SMA), Huntington's disease, deleterious effect
due to excesses of glutamate released as a result of cerebral accidents of traumatic
or other vascular origin will be mentioned.
[0047] In the above disclosed embodiments of the invention, most of the compositions and
methods were related to the aglycone form of said substrates. Alternatively, the invention
further relates to the use of at least one glucuronidated substrate of an UDP-glucuronosyltransferase
(UGT) and salts thereof, for the preparation of a pharmaceutical composition having
an inhibitory effect on the extracellular neuromediator release into an individual
treated with said composition. According to said embodiment, the substrate of an UDP-glucuronosyltransferase
(UGT) is understood as being glucuronidated before its administration to the patient
in need thereof. Said glucuronidation can be obtained by modifying the aglycone substrate
either by chemical or enzymatic way. Chemical glucuronidation can be obtained by using
standard chemical method well known by those in the art consisting for example in
adding a glucuronide group to the aglycone substrate intended to be used (Lacy and
Sainsbury, 1995, Tetrahedron lett., 36,3949-50). Alternatively, the glucuronidated
substrate can be obtained by enzymatic glucuronidation using at least one UDP-glucuronosyltransferase
(UGT) able to glucuronidate said aglycone substrate. In preferred embodiment said
UDP-glucuronosyltransferase (UGT) is naturally expressed in brain, e.g. UDP-glucuronosyltransferase
(UGT) 1A6 or UDP-glucuronosyltransferase (UGT) 2B7.
[0048] According to one embodiment, said glucuronidated UGT substrate is selected among
the group consisting of planar and small glucuronidated phenols, polycyclic aromatic
glucuronidated hydrocarbons, and compounds which are structurally related. More specifically,
said glucuronidated substrate contains at least one moiety selected in the group consisting
in hydroxyl (alcoholic, phenolic, etc...), carboxyl, sulfuryl, carbonyl and amino
(primary, secondary or tertiary) moieties linked to glucuronic acid moiety.
[0049] According to a specific embodiment, said glucuronidated substrate is selected in
the group consisting of 1-Naphthol, 2-Naphthol , 4-nitrophenol , methylsalicylate,
ketoprofen , naproxen, 5-OH tryptamine/serotonin, carprofen/rimadyl, acetaminophen/paracetamol,benzidine,
4-methylumbelliferone (4-MU), silymarin (Venketaramanan et al., 2000, Drug Metab Dispos.,
28,1270-3 ; Silymarin is a mixture of toxifolin, silichristin, silidianin, silybin
A et B, isosilybin A et B) and is further glucuronidated.
[0050] According to another embodiment, said glucuronidated substrate is selected in the
group consisting of transretinoic acid, ASA , AZT , benoxaprofen, benzidine, (benzo(a)pyrene
mbs), buprenorphine, carprofen, chloramphenicol, clofibric acid, flavenoids quercetin,
kaempfenol, cyclosporin, DMXAA, diclofenac, dihydrocodeine DHC, dihydromorphone ,
mefenamic acid, fenemate NSAID, mycophenolic acid MCPA mofetil, fenoprofen, hydromorphone,
ibuprofen, ketoprofen, linoleic acid, lorazepam, losartan, menthol, morphine 3 and
morphine 6, nalbufene, nalmefene, naltrindole, nalorphine, naloxone, naltrexone, S-naproxen,
norcodeine, normorphone, oxycodone, oxymorphone, pirprofen, propanolol, S-oxazepam,
tacrolimus, temazepam, tolcapone, tiaprofenic, valproate, zomepirac, 5-OH tryptamine
and is further glucuronidated.
[0051] The addition salts of the glucuronidated substrates of the invention comprise conventional
salt formed from inorganic or organic acids or bases, such as hydrochloric, hydrobromic,
sulfuric, phosphoric, nitric, perchloric, fumaric, acetic, sodium, lithium, potassium,
magnesium, aluminium, calcium, zinc, ethylenediamine ; formic, benzoic, maleic, tartaric,
citric, oxalic, aspartic acid, and alkane-sulfonic acids is even mentioned.
[0052] The present invention further relates to a method for treating and/or preventing
neuromediator-evoked cytotoxicity in a patient in need thereof comprising administering
to said patient a composition containing a therapeutically effective amount of at
least one glucuronidated substrate of an UDP-glucuronosyltransferase (UGT) as defined
above and a pharmaceutically acceptable carrier.
[0053] The present invention further relates to a method for treating and/or preventing
glutamate-evoked cytotoxicity in a patient in need thereof comprising administering
to said patient a composition containing a therapeutically effective amount of at
least one glucuronidated substrate of an UDP-glucuronosyltransferase (UGT) as defined
above and a pharmaceutically acceptable carrier.
[0054] In a particularly preferred embodiment, said "neuromediator-evoked cytotoxicity"
is a neuromediator-evoked neurotoxicity, and even more preferably a neurodegeneration
(i.e. degeneration of nervous cells).
[0055] In another particularly preferred embodiment, said "glutamate-evoked cytotoxicity"
is a glutamate-evoked neurotoxicity, and even more preferably a neurodegeneration
(i.e. degeneration of nervous cells).
[0056] The present invention further relates to a method for modulating the release of at
least one neuromediator in a patient comprising administering to said patient a composition
containing a therapeutically effective amount of at least one glucuronidated substrate
of an UDP-glucuronosyltransferase (UGT) and a pharmaceutically acceptable carrier,
wherein said substrate is as detailed above.
[0057] The present invention further relates to a method for modulating the release of glutamate
in a patient comprising administering to said patient a composition containing a therapeutically
effective amount of at least one glucuronidated substrate of an UDP-glucuronosyltransferase
(UGT) and a pharmaceutically acceptable carrier, wherein said substrate is as detailed
above.
[0058] The present invention further relates to a method for treating and/or preventing
disease and/or condition associated with the excessive release of at least one neuromediator
in a patient comprising administration to said patient of a composition containing
a therapeutically effective amount of at least one glucuronidated substrate of an
UDP-glucuronosyltransferase (UGT) and a pharmaceutically acceptable carrier, wherein
said substrate is as detailed above.
[0059] The present invention further relates to a method for treating and/or preventing
disease and/or condition associated with the excessive release of glutamate in a patient
comprising administration to said patient of a composition containing a therapeutically
effective amount of at least one glucuronidated substrate of an UDP-glucuronosyltransferase
(UGT) and a pharmaceutically acceptable carrier, wherein said substrate is as detailed
above.
[0060] The UGT substrates (including aglycone or glucuronidated form) described in the present
invention are administered as a composition containing at least one active compound
and a pharmaceutically acceptable carrier. In preparing such a composition, any conventional
pharmaceutically acceptable carrier can be utilized. The carrier material can be an
organic or inorganic inert carrier material suitable for the selected route of administration.
Suitable carriers include water, gelatin, gum arabic, lactose, starch, magnesium stearate,
talc, vegetable oils, polyalkylene-glycols, petroleum jelly and the like. Furthermore,
the pharmaceutical composition may contain other pharmaceutically active agents. Additional
additives such as flavoring agents, preservatives, stabilizers, emulsifying agents,
salts for varying the osmotic pressure, buffers and the like may be added in accordance
with accepted practices of pharmaceutical compounding. Any conventional form such
as tablets, capsules, pills, powders, granules, and the like may be used. Advantageously,
they are in the form of tablets, sugar coated tablets, hard gelatin capsules, capsules,
granules, for oral administration, or solutions or suspensions for administration
via an injectable channel.
[0061] The methods of the invention may be carried out by administering the composition
containing substrates (including aglycone or glucuronidated form) of the invention
by any route whereby drugs are conventionally administered. Such routes include systemic
and local routes. Examples are intravenous, intramuscular, subcutaneous, intracranial,
intraventricular, inhalation (Gradinaru et al., 1999 supra), intraperitoneal, as well
as oral routes. Preferably, the method of the invention is carried out via oral or
intravenous routes of administration.
[0062] In accordance with this invention, the substrates described herein (including aglycone
or glucuronidated form) are useful in pharmaceutically acceptable oral modes. A preferred
oral dosage form comprises tablets, capsules of hard or soft gelatin, methylcellulose
or of another suitable material easily dissolved in the digestive tract. The oral
dosages contemplated in accordance with the present invention will vary in accordance
with the needs of the individual patient as determined by the prescribing physician.
The preferred oral dosage form is capsules or tablets containing from 50 to 500 mg
of a substrate of the invention.
[0063] Typical preparations for intravenous administration would be sterile aqueous solutions
including water/buffered solutions. Intraveneous vehicles include fluid, nutrient
and electrolyte replenishers. Preservatives and other additives may also be present
such as antibiotics and antioxidants. Compositions for bolus i.v. administration may
contain up to 10 mg/ml (10,000 mg/liter) of substrate described herein. Compositions
for i.v. administration preferably contain from about 50 mg/liter to about 500 mg/liter
of at least one substrate described herein.
[0064] In carrying out the method of the invention, substrate of the invention (including
aglycone or glucuronidated form) is generally given to adults daily, preferably orally
or intravenously, in an amount of from about 5 mg/kg to about 30 mg/kg daily, in single
or divided doses, preferably from about 13 mg/kg to about 17 mg/kg daily, with the
precise dosage being varied depending upon the needs of the patient. The doses will
be adapted according to the patient and the pathology to be treated and are for example
1 mg-100 mg/day. In general, this therapy is carried out for a period of about three
months. Alternatively, the method of the invention may be carried out prophylactically
for an indefinite time in those patients who are have a high risk of suffering an
acute neurotoxic event, such as a stroke. For the treatment of an acute neurotoxic
event, the patient should be treated in accordance with the method of the invention
as soon as possible after the diagnosis of the acute neurotoxic event, preferably
within twelve hours, and most preferably within six hours, of the onset of the neurotoxic
event. When the drug is administered orally, it is generally administered at regular
intervals.
[0065] These drugs are notably administered orally or via an injectable channel.
[0066] According to one special embodiment, the UGT substrates (including aglycone or glucuronidated
form) described in the present invention are modified with specific groups facilitating
the passage of said UGT substrate throughout the blood-brain barrier (BBB). One strategy
is lipidization, which involves the addition of lipid-like groups through modification
of the hydrophilic part of the substrate structure. The resulting lipid-soluble substrates
are transported through the BBB by accessing pores that transiently form within the
lipid bilayer. Another strategy is the addition of hydrophobic groups to enhance BBB
transfer by passive diffusion. For examples, addition of methyl groups or acetylation
of amine groups increase lipophilicity and brain penetration. A further approach is
to modify the substrate so that the compound has a structure that mimics a nutrient,
thus giving it access to one of the specialized carrier-mediated transport systems
within the BBB such as amino acid, hexose, vitamin and neuropeptide carriers. An alternative
approach is the coupling of the substrates, via chemical linkers, to small synthetic
peptide-vectors that cross the cellular membranes efficiently such as Pegelin or Penetratin
(for general revue, see Temsamani et al., 2000, Pharm. Sci. Technol. Today, 3, 155-162).
[0067] According to another embodiment, the substrate or preparation of the invention (including
aglycone or glucuronidated form) is administered to the patient in need in combination
with a second compound or formulation aimed at facilitating the transfer throughout
the BBB. For examples, the substrate or preparation of the invention is combined witha
hypertonic solution, a biologically active agents such as bradykinin and angiotensin
peptides, a vasoactive substance such as histamine, leukotrienes which have the ability
of disrupting BBB transiently. According to the present invention, "combination" or
"combined with" means that the substrate or preparation of the invention and the second
compound or formulation aimed at facilitating the transfer throughout the BBB can
be used simultaneously or consecutively or so as to be staggered over time. Simultaneously
refers to a coadministration. In this case, these two essential components can be
mixed to form a composition prior to being administered, or can be administered at
the same time to the patient in need. It is also possible to administer them consecutively,
that is to say one after the other, irrespective of which component is administered
first. Finally, it is possible to use a mode of administration which is staggered
over time or is intermittent and which stops and restarts at intervals which may or
may not be regular. It is pointed out that the routes and sites of administration
of the two components can be different. The time interval between the administration
and the routes and sites of administration can be defined by the skilled person. It
is possible to recommend an interval of from 10 min to 72 h, advantageously of from
30 min to 48 h, preferably of from 1 to 24 h and, very preferably, of from 1 to 6
h.
[0068] Pharmacogenomic analysis (see for example Ciotti et al., 1997, Pahrmacogenetics,
7, 485-495) have shown that polymorphisms in the UGT genes can result in affected
enzyme activity and in lowering or increasing the patient reactivity towards UGT substrates.
For example, missense mutations in the UGT1A6 have been identified (A541→G541 and
A552→C552) which cause variation in the UGT1A6 activity and therefore variation in
the rate of their substrate metabolization (for more detail, see Ciotti et al., 1997,
Pharmacogenetics, 7, 485-495). Thus, according to another special embodiment, the
treatment and/or prevention methods of the present invention further comprise a preliminary
step consisting in establishing if the patient to be treated is presenting a genetic
polymorphism resulting in less or improved UGT activity. More specifically, said preliminary
step consists in determining by nucleotide sequence analysis of the patient genomic
DNA if the UGT1A6 gene of said patient to be treated comprises at least one of the
mutation A541→G541 (T181A mutation) or A552→C552 (R184S mutation). Details of said
method are disclosed for example in Ciotti et al., 1997, Pharmacogenetics, 7, 485-495
the full content of which is incorporated hereby by reference.
[0069] All publications and patent applications cited in this specification are herein incorporated
by reference as if each individual publication or patent application were specifically
and individually indicated to be incorporated by reference. Although the foregoing
invention has been described in some detail by way of illustration and example for
purposes of clarity of understanding, it will be readily apparent to those of ordinary
skill in the light of the teachings of this invention that certain changes and modifications
may be made thereto without departing from the spirit or scope of the appended claims.
[0070] The invention has been described in an illustrative manner, and it is to be understood
that the terminology which has been used is intended to be in the nature of words
of description rather than of limitation. Obviously, many modifications and variations
of the present invention are possible in light of the above teachings. It is therefore
to be understood that within the scope of the appended claims, the invention may be
practised otherwise than as specifically described. Accordingly, those skilled in
the art will recognize, or able to ascertain using no more than routine experimentation,
many equivalents to the specific embodiments of the invention described specifically
herein. Such equivalents are intended to be encompassed in the scope of the following
claims.
[0071] These and other embodiments are disclosed or are obvious from and encompassed by
the description and examples of the present invention. Further literature concerning
any one of the methods, uses and compounds to be employed in accordance with the present
invention may be retrieved from public libraries, using for example electronic devices.
For example the public database "Medline" may be utilized which is available on Internet,
e.g. under http://www.ncbi.nlm.nih.gov/PubMed/medline.html. Further databases and
addresses, such as http://www.ncbi.nlm.nih.gov/, http://www.infobiogen.fr/, http://www.fmi.ch/biology/resea
rch_tools.html, http://www.tigr.org/, are known to the person skilled in the art and
can also be obtained using, e.g., http://www.lycos.com. An overview of patent information
in biotechnology and a survey of relevant sources of patent information useful for
retrospective searching and for current awareness are given in Berks, TIBTECH 12 (1994),
352-364.
EXAMPLES
[0072] Primary cortical neuron cultures. Cortices from foetal Wistar rats (E16) were dissected and maintained in PBS without
calcium and magnesium. They were then treated in PBS containing 0.25% Trypsin for
15 min at 37°C and placed in a complete medium containing Horse Serum for inhibiting
trypsin activity. The cortices were then dissociated in complete medium (Neurobasal,
2% Horse Serum, 2mM Glutamine, B27 supplement (1X), 100µg/ml Gentamicyn, 10µM β Mercaptoethanol).
Cells were seeded at 10
5 cells per well in a 96 well plate previously coated with Poly-Ornithine at 9µg/ml.
The cultures were maintained at 37°C in a humidified incubator with 5% CO
2. At day 2, 5µM Cytosine Arabinofuranoside was added to the cell cultures in order
to inhibit glial cell proliferation. At day 7, the primary cultures of neurons were
available for further experiments.
[0073] Assessment of neuron cell death. Neuron cell death was estimated by measuring the release of the cytosolic enzyme,
lactate dehydrogenase (LDH), into the medium of cell cultures. The LDH release quantification
was performed using the colorimetric CytoTox96® nonradioactive assay (Promega). Briefly,
100µL of cell culture medium were removed and centrifuged for 4 min at 1500 rpm in
order to remove cellular debris. 50µL were then added to 50µL of assay buffer and
placed in the dark for 30min at room temperature. The reaction was stopped with 50µL
of stop solution and the absorbance at 490nm was measured. The percentage of cell
death was calculated by comparison with the control.
[0074] Assessment of glutamate release. Glutamate released by neurons in culture was evaluated in aliquots of the supernatants
by means of a chemiluminescent enzymatic assay (Israel et al., Neurochem Int. 1993
Jan;22(1):53-8).
EXAMPLE 1
Neuroprotection assay.
[0075] Diazepam (Sigma) and the glucuronide derivative of Diazepam, named Temazepam (Alltech),
a substrate of the UDP-glucuronosyltransferase 2B7 (UGT2B7), were used to illustrate
the preventing and/or treating effects of UGT2B7 substrates on glutamate-induced neurological
disorders.
[0076] Diazepam and Temazepam were used to protect primary cultures of neurons, isolated
from the rat brain cortex, from neurotoxin-induced cell death.
[0077] The neurons were treated
in vitro with 3-nitropropionic acid (3-NP, Sigma) at 400µM, a toxin which induces epileptic
seizures in animal models, in absence or presence of Diazepam and Temazepam at 0.1,
1 and 5µM. After 24 hours, the cultures of cells were analysed for the level of toxin-evoked
cell death by means of lactate dehydrogenase release quantification.
[0078] The results (Figure 2) show that Diazepam protects neurons from 3-NP at all tested
doses but that Temazepam significantly reinforces said cell protection by about 50%
at 1 and 5µM. These data indicate that glucuronide modification of Diazepam influences
cell survival upon exposure to a toxic compound.
[0079] Similar results were obtained with another neurotoxin, the kainic acid at 100µM (Fisher
Bioblock), which mimics glutamate-induced epileptic seizures in animal models. A better
neuroprotection was observed with 1 and 5µM of Temazepam compared with Diazepam at
the same concentrations (Figure 3).
EXAMPLE 2
Neuroprotection assay.
[0080] 1-Naphthol (Merck), a substrate of UDP-glucuronosyltransferase 1A6 (UGT1A6), was
used to illustrate the preventing and/or treating effects of UGT1A6 substrates on
glutamate-induced neurological disorders.
[0081] 1-Naphthol was used to protect primary cultures of neurons, isolated from the rat
brain cortex, from neurotoxin-induced cell death.
[0082] The neurons were treated
in vitro with 5µM of Ionomycin (Sigma), a calcium ionophore, in presence of 10µM of 1-Naphthol,
in absence or in presence of 5 or 10µM of 3-methylcholanthrene (3-MC, Aldrich), an
UGT inducer which allows the
in situ glucuronidation of 1-Naphthol.
[0083] The results (Figure 4) show that Naphthol protects neurons from Ionomycin-evoked
cell death and that glucuronidation of 1-Naphthol by 3-MC reinforces, dose dependently,
the protective effect of 1-Naphthol.
[0084] These data indicate that glucuronide derivatives of UGT1A6 substrates present better
neuroprotective effects than the parent molecules.
Glutamate level determination.
[0085] The glutamate released by neurons was followed by an enzymatic assay coupled to a
chemiluminescent reaction. In this experiment, primary cultures of neurons isolated
from the rat brain cortex, were treated by 1-Naphthol at 10µM with or without 3-MC
at 1 or 5µM. The results (Figure 5) indicate that 1-Naphthol reduces glutamate release
and that glucuronidation of 1-Naphthol by 3-MC reinforces, dose dependently, the decrease
of glutamate.
[0086] This experiment indicates that the protective effect of glucuronidation on UGT1A6
substrates is associated with an improvement of the decrease of glutamate release.
Figure 1 (A/B) : Illustrates the chemical structures of designed compounds cited throughout
the specification.
Figure 2 : Effect of Diazepam and Temazepam on neuron cell death. Neuron cell death
in culture was arbitrarily set at 0. The cells were treated with the vehicle (lane
1) and with 3-NP at 400µM alone (lane 2) or simultaneously with one of the three tested
concentrations of Diazepam (lanes 3-5) and Temazepam (lanes 6-8). Results indicate
that Temazepam at 1µM (lane 7) and 5µM (lane 8) but not at 0.1µM (lane 6) improves
the neuroprotective effect of Diazepam observed at all concentrations (lanes 3-5).
Figure 3 : Effect of Diazepam and Temazepam on neuron cell death. Neuron cell death
in culture was arbitrarily set at 0. The cells were treated with the vehicle (lane
1) and with kainic acid at 100µM alone (lane 2) or simultaneously with one of the
three tested concentrations of Diazepam (lanes 3-5) and Temazepam (lanes 6-8). Results
indicate that Temazepam at 1µM (lane 7) and 5µM (lane 8) but not at 0.1µM (lane 6)
improves the neuroprotective effect of Diazepam observed at all concentrations (lanes
3-5).
Figure 4 : Effect of 1-Naphthol and 1-Naphthol glucuronide on neuron cell death. Neuron
cell death in culture was arbitrarily set at 0. The cells were treated with the vehicle
(lane 1) and with Ionomycin at 5µM alone (lane 4) or simultaneously with 1-Naphtol
at 10µM (lane 7) with 3-Methylcholanthrene at 5µM (lane 8) or 10µM (lane 9). The two
tested doses of 3-Methylcholanthrene were evaluated in absence (lanes 2, 3) or presence
(lanes 5, 6) of Ionomycin and considered as controls. Results indicate that 3-Methylcholanthrene
alone has no effect on the natural death of neurons (lanes 2, 3) and that it has no
protective effect against the toxicity induced by Ionomycin (lanes 5, 6). Results
also show that the protection induced by 1-Naphthol at 10µM (lane 7) is improved by
the glucuronidation induced by 3-Methylcholanthrene at 5µM (lane 8) and 10µM (lane
9).
Figure 5 : Effect of 1-Naphthol and 1-Naphthol glucuronide on glutamate released by
neurons in culture. Glutamate release was evaluated by means of a chemiluminescent
enzymatic assay. The basal levels of glutamate release were quantified in the non-treated
cells (lane 1), in cells treated with the vehicle (lane 2) and in cells treated with
3-Methylcholanthrene at the highest dose tested in this experiment, 5µM (lane 3).
The effect of 1-Naphthol at 1µM (lane 4) and 10µM (lane 7) was evaluated in presence
of 3-Methylcholanthrene at 1µM (lanes 5, 8) and at 5µM (lanes 6, 9). Results indicate
that the vehicle and the glucuronidation inducer 3-Methylcholanthrene have no effect
on glutamate release (lanes 2, 3) whereas 1-Naphthol at both concentrations reduces
the release of glutamate (lanes 4, 7). In addition, the glucuronidation of 1-Naphthol
at both doses improves the decrease of glutamate for the two tested doses of 3-Methylcholanthrene
(lanes 5, 6, 8, 9). This improvement becomes statistically significant at the highest
dose of 3-Methylcholanthrene (lanes 6, 9) whatever the 1-Naphthol dose is.
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1. Use of at least one substrate of an UDP-glucuronosyltransferase (UGT), and salts thereof,
for the preparation of a pharmaceutical composition having an inhibitory effect on
the extracellular glutamate release into an individual treated with said composition.
2. The use of claim 1, wherein said UDP-glucuronosyltransferase (UGT) is expressed in
brain and is selected among the group consisting of UDP-glucuronosyltransferase 1A6
(UGT1A6) and UDP-glucuronosyltransferase 2B7 (UGT2B7).
3. The use of claim 1 or 2, wherein said UGT substrate is selected among the group consisting
of planar and small phenols, polycyclic aromatic hydrocarbons, and compounds which
are structurally related.
4. The use of claims 1-3, wherein said substrate is a substrate of the UDP-glucuronosyltransferase
1A6 (UGT1A6) and is selected in the group consisting of 1-Naphthol, 2-Naphthol , 4-nitrophenol
, methylsalicylate, ketoprofen , naproxen, 5-OH tryptamine/serotonin, carprofen/rimadyl
, acetaminophen/paracetamol ,benzidine, 4-methylumbelliferone (4-MU), silymarin.
5. The use of claims 1-3, wherein said substrate is a substrate of the UDP-glucuronosyltransferase
2B7 (UGT2B7) and is selected in the group consisting of transretinoic acid, ASA ,
AZT , benoxaprofen , benzidine, (benzo(a)pyrene mbs), buprenorphine, carprofen, chloramphenicol,
clofibric acid , flavenoids quercetin, kaempfenol, cyclosporin, DMXAA, diclofenac,
dihydrocodeine DHC, dihydromorphone, mefenamic acid, fenemate NSAID, mycophenolic
acid MCPA mofetil, fenoprofen, hydromorphone, ibuprofen, ketoprofen, linoleic acid,
lorazepam, losartan, menthol, morphine 3 and morphine 6, nalbufene, nalmefene, naltrindole,
nalorphine, naloxone, naltrexone, S-naproxen, norcodeine, normorphone, oxycodone,
oxymorphone, pirprofen, propanolol, S-oxazepam, tacrolimus, temazepam, tolcapone,
tiaprofenic, valproate, zomepirac, 5-OH tryptamine.
6. The use of claims 1-5, wherein said pharmaceutical composition is intended for treating
and/or preventing glutamate-evoked cytotoxicity in a patient in need.
7. The use of claims 1-5, wherein said pharmaceutical composition is intended for treating
and/or preventing glutamate-evoked neurotoxicity.
8. The use of claims 1-5, wherein said pharmaceutical composition is intended for treating
and/or preventing neurodegeneration.
9. The use of claims 1-5, wherein said pharmaceutical composition is intended for modulating
the release of glutamate in a patient.
10. The use of claims 1-5, wherein said pharmaceutical composition is intended for treating
and/or preventing disease and/or condition associated with the excessive release of
glutamate in a patient.
11. The use of claim 10, wherein said disease and/or condition associated with the excessive
release of glutamate is selected among the group consisting of epileptic seizures,
acute and chronic neurodegenerative diseases, ischemia, Alzheimer's, Huntington's,
Parkinson's diseases, multiple sclerosis (MS), amyotrophic lateral sclerosis (ALS),
spinal muscular atrophy (SMA), retinopathy, stroke and traumatic brain injury, drug-induced
neurotoxicity, pain, hormonal balance, blood pressure, thermoregulation, respiration,
learning, pattern recognition, memory, and disorders subsequent to hypoxia or hypoglycaemia.
12. Use of at least one glucuronidated substrate of an UDP-glucuronosyltransferase (UGT)
and salts thereof, for the preparation of a pharmaceutical composition having an inhibitory
effect on the extracellular glutamate release into an individual treated with said
composition.
13. The use of claim 12, wherein said glucuronidated UGT substrate is selected among the
group consisting of planar and small glucuronidated phenols, polycyclic aromatic glucuronidated
hydrocarbons, and compounds which are structurally related.
14. The use of claims 12-13, wherein said glucuronidated substrate is selected in the
group consisting of 1-Naphthol, 2-Naphthol , 4-nitrophenol , methylsalicylate, ketoprofen
, naproxen, 5-OH tryptamine/serotonin, carprofen/rimadyl , acetaminophen/paracetamol
,benzidine, 4-methylumbelliferone (4-MU), silymarin and is further glucuronidated.
15. The use of claims 12-13, wherein said glucuronidated substrate is selected in the
group consisting of transretinoic acid, ASA , AZT , benoxaprofen , benzidine, (benzo(a)pyrene
mbs), buprenorphine, carprofen, chloramphenicol, clofibric acid , flavenoids quercetin,
kaempfenol, cyclosporin, DMXAA, diclofenac, dihydrocodeine DHC, dihydromorphone, mefenamic
acid, fenemate NSAID, mycophenolic acid MCPA mofetil, fenoprofen, hydromorphone, ibuprofen,
ketoprofen, linoleic acid , lorazepam, losartan, menthol, morphine 3 and morphine
6, nalbufene, nalmefene, naltrindole, nalorphine, naloxone, naltrexone, S-naproxen,
norcodeine, normorphone, oxycodone, oxymorphone, pirprofen, propanolol, S-oxazepam,
tacrolimus, temazepam, tolcapone, tiaprofenic, valproate, zomepirac, 5-OH tryptamine
and is further glucuronidated.